Figure 3
Morphology and function of Ncadk.i.;p53 mammary glands are comparable to control samples. (A) Whole mount carmine staining of Ncadk.i.;p53 mammary glands showed comparable lobulo-alveolar structures to control;p53, while Ecadk.o.;p53 mammary glands possess only poorly developed structures. This observation was confirmed by H&E-stained paraffin sections showing intact alveoli in Ncadk.i.;p53 and control;p53 mammary glands and collapsed lobulo-alveolar structures in Ecadk.o.;p53 mammary glands (arrowheads). The accumulation of fibrous tissue surrounding the alveoli in Ncadk.i.;p53 was still present (arrows). Scale bar: Carmin: 3 mm; H&E: 100 μm (B) Immunofluorescence on paraffin sections revealed basolateral localization of N-cad in the alveoli of Ncadk.i.p53 mammary glands, resembling E-cad staining in control;p53 samples. DAPI was used to visualize the nuclei in Ecadk.o.;p53 sections, which were negative for E-cad. The detection of WAP was used to verify the function of the alveoli. Ncadk.i.p53 and control;p53 showed similar WAP levels, while Ecadk.o.p53 exhibited highly decreased levels. Scale bar: 100 μm (C) Western Blot analysis of Ncadk.i.p53, Ecadk.o.;p53, and control;p53 mammary gland lysates. Notably, Ncadk.i.p53 shows a constitutive activation of p-Stat3. control;p53, WAP::Cre; Ecadfl/+; DAPI, 4',6-diamidino-2-phenylindole; E-cad, E-cadherin; Ecadk.o.;p53, WAP::Cre;Ecadfl/fl;p53fl/+; H&E, Hematoxylin and Eosin; N-cad, N-cadherin; Ncadk.i.;p53, WAP::Cre;EcadNcad/fl;p53fl/+; p53fl/+; WAP, Whey acidic protein.